Detection of enterovirus viraemia in blood donors

Welch, Jonathan; McGowan, Karen; Searle, Benjamin; Gillon, J.; Jarvis, Lisa; Simmonds, Peter

doi:10.1046/j.1423-0410.2001.00035.x

Cited by 22 publications

(27 citation statements)

References 11 publications

(9 reference statements)

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“…Conserved regions between HRV-A, -B, and -C coincided with those of HEV-A to HEV-D, enabling a combined HRV-HEV amplification method, although the incorporation of HEV and HRV-C amplification necessitated the use of separate inner sense primers; for example, HEV 5Ј UTR sequences differed from HRV at several positions in the primer-binding target ( Table 1; the bases that are different are underlined and in boldface). The sensitivity of the combined HEV-HRV PCR was compared to that of a previously described 5Ј UTR-based nested PCR previously used for the screening of clinical specimens (cerebrospinal fluid in suspected viral meningitis/encephalitis cases [19]) and surveillance samples (blood donor screening [39]) ( Table 2). On the basis of the proportion of replicates positive at each dilution, the two assays showed identical sensitivities.…”

Section: Resultsmentioning

confidence: 99%

Screening Respiratory Samples for Detection of Human Rhinoviruses (HRVs) and Enteroviruses: Comprehensive VP4-VP2 Typing Reveals High Incidence and Genetic Diversity of HRV Species C

et al. 2009

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Rhinovirus infections are the most common cause of viral illness in humans, and there is increasing evidence of their etiological role in severe acute respiratory tract infections (ARTIs). Human rhinoviruses (HRVs) are classified into two species, species A and B, which contain over 100 serotypes, and a recently discovered genetically heterogeneous third species (HRV species C). To investigate their diversity and population turnover, screening for the detection and the genetic characterization of HRV variants in diagnostic respiratory samples was performed by using nested primers for the efficient amplification of the VP4-VP2 region of HRV (and enterovirus) species and serotype identification. HRV species A, B, and C variants were detected in 14%, 1.8%, and 6.8%, respectively, of 456 diagnostic respiratory samples from 345 subjects (6 samples also contained enteroviruses), predominantly among children under age 10 years. HRV species A and B variants were remarkably heterogeneous, with 22 and 6 different serotypes, respectively, detected among 73 positive samples. Similarly, by using a pairwise distance threshold of 0.1, species C variants occurring worldwide were provisionally assigned to 47 different types, of which 15 were present among samples from Edinburgh, United Kingdom. There was a rapid turnover of variants, with only 5 of 43 serotypes detected during both sampling periods. By using divergence thresholds and phylogenetic analysis, several species A and C variants could provisionally be assigned to new types. An initial investigation of the clinical differences between rhinovirus species found HRV species C to be nearly twice as frequently associated with ARTIs than other rhinovirus species, which matches the frequencies of detection of respiratory syncytial virus. The study demonstrates the extraordinary genetic diversity of HRVs, their rapid population turnover, and their extensive involvement in childhood respiratory disease.

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Section: Resultsmentioning

confidence: 99%

Screening Respiratory Samples for Detection of Human Rhinoviruses (HRVs) and Enteroviruses: Comprehensive VP4-VP2 Typing Reveals High Incidence and Genetic Diversity of HRV Species C

et al. 2009

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“…Scottish blood donor samples were obtained in a previous study, in which a highly sensitive reverse transcriptase PCR (RT-PCR) amplification method using highly conserved primers specific for the 5Ј UTR was used to screen approximately 330,000 blood donation samples in pools of 95 between 2000 and 2001 (44). Viremic donors were identified by testing cross pools and individual donations; through the use of this procedure, no pools were found that contained more than one PCR-positive component donation (44). Samples found to be positive using the 5Ј UTR RT-PCR assay were further investigated by amplification and sequencing of regions of the capsid (VP1) for serotype identification as previously described (44).…”

Section: Methodsmentioning

confidence: 99%

“…Viremic donors were identified by testing cross pools and individual donations; through the use of this procedure, no pools were found that contained more than one PCR-positive component donation (44). Samples found to be positive using the 5Ј UTR RT-PCR assay were further investigated by amplification and sequencing of regions of the capsid (VP1) for serotype identification as previously described (44). The sequence of the 251-nt region of VP1 (positions 3090 to 3341, numbered relative to the poliovirus P3/Leon/37 strain; accession number K01392) was capable of reliably identifying the enterovirus serotype in each of the blood donor samples.…”

Section: Methodsmentioning

confidence: 99%

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Frequency and Dynamics of Recombination within Different Species of Human Enteroviruses

Simmonds

Welch

2006

J Virol

239

230

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Enteroviruses are members of the family Picornaviridae that cause widespread infections in human and other mammalian populations. Enteroviruses are genetically and antigenically highly variable, and recombination within and between serotypes contributes to their genetic diversity. To investigate the dynamics of the recombination process, sequence phylogenies between three regions of the genome (VP4, VP1, and 3Dpol) were compared among species A and B enterovirus variants detected in a human population-based survey in Scotland between 2000 and 2001, along with contemporary virus isolates collected in the same geographical region. This analysis used novel bioinformatic methods to quantify phylogenetic compatibility and correlations with serotype assignments of evolutionary trees constructed for different regions of the enterovirus genome. Species B enteroviruses showed much more frequent, time-correlated recombination events than those found for species A, despite the equivalence in population sampling, concordant with a linkage analysis of previously characterized enterovirus sequences obtained over longer collection periods. An analysis of recombination among complete genome sequences by computation of a phylogenetic compatibility matrix (PCM) demonstrated sharply defined boundaries between the VP2/VP3/VP1 block and sequences to either side in phylogenetic compatibility. The PCM also revealed equivalent or frequently greater degrees of incompatibility between different parts within the nonstructural region (2A-3D), indicating the occurrence of extensive recombination events in the past evolution of this part of the genome. Together, these findings provide new insights into the dynamics of species A and B enterovirus recombination and evolution and into the contribution of structured sampling to documenting reservoirs, emergence, and spread of novel recombinant forms in human populations.The genus Enterovirus in the family Picornaviridae is a group of nonenveloped RNA viruses that cause a wide range of diseases in humans and other mammals. The enterovirus genome is a single strand of positive-sense RNA of approximately 7,500 nucleotides (nt) comprising a long open reading frame flanked 5Ј and 3Ј by untranslated regions (UTRs). The encoded polyprotein is subsequently cleaved to produce structural (capsid proteins VP1 to VP4) and nonstructural (2A and 3D) proteins. Primary infection with an enterovirus leads to viral replication in the tissue around the gastrointestinal tract, followed by a transient viremia and sometimes migration into other tissues (2, 34). Although infection in immunocompetent individuals is often asymptomatic or causes mild febrile illness, a persistent or widely disseminated, systemic infection associated with severe disease outcomes is observed in highly immunosuppressed individuals and neonates (6, 13, 18).Enteroviruses were originally classified as polioviruses, coxsackie type A or B viruses (CVA and CVB), or echoviruses (enteric cytopathic human orphan viruses), depending on the infectious...

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“…3 and 4). Targeted amplification from stool samples, using pan-HEV primers (39), indicated that 23 of these 35 tested patients were positive for HEV infection (20), 17 of which were also identified using viral metagenomics. Other known enteric viruses observed included adenovirus, picobirnavirus, rotavirus, Aichi virus, parechovirus, and rhinovirus, as well as assorted plant viruses of the families Partitiviridae and Tobamoviridae ( Fig.…”

Section: Samplingmentioning

confidence: 99%

Metagenomic Analyses of Viruses in Stool Samples from Children with Acute Flaccid Paralysis

Victoria

Kapoor

et al. 2009

J Virol

342

345

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We analyzed viral nucleic acids in stool samples collected from 35 South Asian children with nonpolio acute flaccid paralysis (AFP). Sequence-independent reverse transcription and PCR amplification of capsid-protected, nuclease-resistant viral nucleic acids were followed by DNA sequencing and sequence similarity searches. Limited Sanger sequencing (35 to 240 subclones per sample) identified an average of 1.4 distinct eukaryotic viruses per sample, while pyrosequencing yielded 2.6 viruses per sample. In addition to bacteriophage and plant viruses, we detected known enteric viruses, including rotavirus, adenovirus, picobirnavirus, and human enterovirus species A (HEV-A) to HEV-C, as well as numerous other members of the Picornaviridae family, including parechovirus, Aichi virus, rhinovirus, and human cardiovirus. The viruses with the most divergent sequences relative to those of previously reported viruses included members of a novel Picornaviridae genus and four new viral species (members of the Dicistroviridae, Nodaviridae, and Circoviridae families and the Bocavirus genus). Samples from six healthy contacts of AFP patients were similarly analyzed and also contained numerous viruses, particularly HEV-C, including a potentially novel Enterovirus genotype. Determining the prevalences and pathogenicities of the novel genotypes, species, genera, and potential new viral families identified in this study in different demographic groups will require further studies with different demographic and patient groups, now facilitated by knowledge of these viral genomes.

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Detection of enterovirus viraemia in blood donors

Cited by 22 publications

References 11 publications

Screening Respiratory Samples for Detection of Human Rhinoviruses (HRVs) and Enteroviruses: Comprehensive VP4-VP2 Typing Reveals High Incidence and Genetic Diversity of HRV Species C

Screening Respiratory Samples for Detection of Human Rhinoviruses (HRVs) and Enteroviruses: Comprehensive VP4-VP2 Typing Reveals High Incidence and Genetic Diversity of HRV Species C

Frequency and Dynamics of Recombination within Different Species of Human Enteroviruses

Metagenomic Analyses of Viruses in Stool Samples from Children with Acute Flaccid Paralysis

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